Evaluation of biochemical and serological methods to identify and clustering yeast cells of oral Candida species by CHROMagar test, SDS-PAGE and ELISA

Abstract

The purpose of this work was to evaluate biochemical and serological methods to characterize and identify Candida species from the oral cavity. The strains used were five Candida species previously identified: C. albicans, C. guilliermondii, C. parapsilosis, C. krusei, C. tropicalis, and Kluyveromyces marxianus, as a negative control. The analyses were conducted through the SDS-PAGE associated with statistical analysis using software, chromogenic medium, and CHROMagar Candida (CA), as a differential medium for the isolation and presumptive identification of clinically important yeasts and an enzyme-linked immunoabsorbent assay (ELISA), using antisera produced against antigens from two C. albicans strains. This method enabled the screening of the three Candida species: C. albicans, C. tropicalis, and C. Krusei, with 100% of specificity. The ELISA using purified immunoglobulin G showed a high level of crossreaction against protein extracts of Candida species. The SDS-PAGE method allowed the clustering of species-specific isolates using the Simple Matching coefficient, S SM = 1.0. The protein profile analysis by SDS-PAGE increases what is known about the taxonomic relationships among oral yeasts. This methodology showed good reproducibility and allows collection of useful information for numerical analysis on information relevant to clinical application, and epidemiological and systematical studies.

Clinical and microbiological diagnosis of oral candidiasis

“…Many enzyme techniques are commercially available (CHROMagar Candida®, Candida ID®, Candida ID 2®, CHROMagar-PAL®, Brilliance® Candida Agar, Colorex®, Chromogen albicans®, Albicans ID2®, Candiselect®, Fluoroplate® and Agar SDCA-MUAG®)(5,7,8,14,22). However, the most frequently used system is CHROMagar Candida®, where C. albicans grows forming smooth green colonies, C. tropicalis forms smooth blue colonies, and C. krusei forms rough pink colonies (3,8,9,26). Another widely used chromogenic method is Candida ID®, where C. albicans grows to form smooth blue colored colonies, C. tropicalis and C. guillermondii generate pink colonies, and the rest of the species appear as white colonies (22) (Fig.…”

“…The host predisposing factors include endocrine alterations (diabetes mellitus, pregnancy, renal failure and hyperthyroidism)(2,3), immune depression (normally associated to antineoplastic treatments or immunosuppression in transplant patients, as well as agammaglobulinemia or cellular immune defects)(1,4-8), acquired immunodeficiency syndrome (AIDS) or hematological and immune disorders such as agranulocytosis (neutropenia). Other predisposing conditions are malignant diseases such as lymphomas or leukemias, aplastic anemia, drug treatments (long-term administration of broad spectrum antibiotics, corticosteroids, antidepressants, antineoplastic drugs and immunosuppressants)(9-11), hyposialia (produced by disorders such as Sjögren’s disease, drugs or radiotherapy), and terminal or end-stage systemic diseases (10,12,13). The oral microenviron-ment-modifying factors in turn include poorly fitting dentures (14-17), loss of vertical dimension, chronic antiseptic use, prolonged dummy use in children, poor oral hygiene (16), smoking and alcoholism (10,18).…”

Antimicrobial potential of some plant extracts against Candida species

“…Although the colonisation by Candida albicans is common and causes severe injuries in immunocompromised patients, other Candida species have been isolated from such patients, healthy patients and children with Down's syndrome, such as C. glabrata, C. krusei, C. tropicalis, C. lusitaniae, C. parapsilosis, C. guilliermondii (Erköse and Erturan, 2007;Höfling et al, 2001;Ribeiro et al, 2006;Rodrigues et al, 2004) and C. dubliniensis. These yeasts were recognised as the source of mucosal infection in HIV-positive patients and regarded as a significant cause of infections in humans, such as abdominal infections and fungemia (Erköse and Erturan, 2007).…”

Histological and immunohistochemical findings of the action of botulinum toxin in salivary gland: systematic review

“…On the other hand, the decrease in salivary flow and xerostomia, the person may develop severe weakness in oral health, difficulty with speech, chewing, swallowing, changes in the mucous membrane, tooth loss (Sanioto et al, 2013), among others as microbial infections such as candidiasis caused by pathogenic species of Candida (Rodrigues et al, 2004).…”